对虾的免疫机制及其疾病预防策略的研究

在对虾的细胞免疫中血细胞是主要的作用因素,而体液免疫是血淋巴中的一些酶和调节因子,机体还可以被诱导产生特殊的免疫保护反应.应用免疫增强剂、疫苗和基因工程技术为预防对虾病害提供了有效的途径.本文根据国内外的有关资料,就对虾的免疫机制和疾病预防策略进行了综述.     

玉米叶片受新月弯孢菌侵染后的细胞病理学变化

 本文利用透射电子显微镜技术与细胞化学技术研究了玉米叶片受弯孢菌侵染后的超微结构和细胞壁的组成成份变化。透射电镜观察发现,病菌侵入后,菌丝先在寄主细胞间扩展,随着寄主细胞病变、坏死,菌丝可进入寄主细胞形成胞内菌丝。随病菌侵入和在寄主体内扩展,寄主细胞先后发生了一系列的超微结构变化,叶绿体、液泡等细胞器解体,出现质壁分离现象,并最终解体、坏死、变形。细胞化学标记定位发现,受侵寄主细胞壁中纤维素、木聚糖和果胶质的标记密度明显低于未接种的健康组织,表明细胞壁降解酶(如纤维素酶、木聚糖酶和果胶酶)的产生与病菌侵染和致病过程密切相关。     

不同光湿环境对葡萄花芽分化和叶绿体类囊体膜磷酸酯酶活性的影响

以欧亚种葡萄美人指(Vitisviniferacv.ManicureFinger)为试材,正常光湿为对照,采用正常光偏高湿、偏弱光正常湿度、偏弱光高湿、偏弱光临界高湿、临界弱光偏高湿和临界弱光临界高湿6种处理。试验表明:单一偏弱光处理,葡萄叶片叶绿素含量上升,其余处理叶绿素含量下降;6种处理叶绿体类囊体膜磷酸酯酶活性都低于对照,叶绿体类囊体膜磷酸酯酶活性与光照和湿度有关;正常光偏高湿和偏弱光正常湿度部分芽能分化花原基,但在偏弱光高湿或偏弱光临界高湿花芽分化质量严重下降,临界弱光偏高湿和临界弱光临界高湿几乎未发现花原基;单一的弱光因子比单一的高湿因子对叶绿体类囊体膜磷酸酯酶活性和花芽形态分化影响更大,但弱光与高湿同时存在比单一弱光或高湿因子作用于葡萄时,叶绿体类囊体膜磷酸酯酶活性下降更为显著,花芽更难以形成。     

钙及其拮抗剂对苹果果肉质膜透性的调节作用

采用培养果肉圆片的方法,研究了Ca2+及其拮抗剂对苹果果肉质膜透性的调节作用。结果表明,CaCl2(1、10mmol/L)降低果肉膜透性和溶质外渗速率(Js);细胞膜Ca2+通道阻塞剂Verapamil(100μmol/L)的影响不显著;细胞外Ca2+螯合剂EGTA(5mmol/L)、CaM的拮抗剂CPZ、TFP(100μmol/L)明显提高果肉膜透性和细胞溶质外渗速率。培养24h时,CaCl2能明显维持较高的SOD活性和ACC向乙烯的转化能力,EGTA、Verapamail、CPZ和TFP的作用相反。这些说明Ca2+对果肉细胞膜具有保护作用,而减少细胞外Ca2+和抑制细胞内Ca2+-CaM功能对果肉细胞膜具有伤害作用。     

Expression of cyclooxygenase-2 in IL-1β-stimulated mesangial cells is medicated by NF-κB/IκB signal pathway

AIM: To investigate the role of NF-κB/IκB signal pathway in the regulation of cyclooxygenase-2 (COX-2) expression in human mesangial cells (HMC). METHODS: The PGE₂ concentration in supernatants of HMC was measured by radioimmunoassay. COX-2 mRNA and protein expression were determined by RT-PCR and Western blot. Electrophoretic mobility shift assay (EMSA) and Western blot were used to detect the activity of NF-κB and degradation of IκB. RESULTS: IL-1β significantly upregulated COX-2 expression and PGE₂ production in HMC. Significant up-regulation of NF-κB activation, nuclear translocation of p65 subunit, and degradation of IκB α and IκB β were observed in IL-1β-induced HMC. CONCLUSION: Expression of COX-2 in IL-1β-induced HMC is mediated by NF-κB/IκB signal pathway.     

Regulation of MMP-2 and TIMP-3 expression by uPA signal transduction system in human bone giant cell tumor

AIM: To study effects of urokinase-type plasminogen activator (uPA) signal transduction on expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) in giant cell tumor of bone (GCT). METHODS: Expression of uPAR, MMP-2 and TIMP-3 in GCT tissue was detected by immunohistochemistry. Phosphorylation level of mitogen-activated protein kinase (p44) in uPA/uPAR signal pathway in cultured GCT cells was detected by immunoprecipitation. The expression of MMP-2 and TIMP-3 in cultured cells after treatment with uPA-ATF or anti-uPAR antibody was also detected by Western blotting. RESULTS: 1) Urokinase-type plasminogen activator receptor (uPAR) was positive on the cell membrane and in cytoplasm of some mononuclear stromal cells (MSCs) and multinucleated giant cells (MGCs); 2) MMP-2 was positive in the cytoplasm and on the cell membrane of almost all of MSCs and some of MGCs. The polar distribution of MMP-2 in the cytoplasm of MGCs was especially obvious; 3) The expression of TIMP-3 of some MSCs and MGCs in GCT was much lower than MMP-2. The positive signal also showed a prominent polarity; 4) After treatment with uPA-ATF, the phosphorylation level of p44 in GCT cultured cells was much higher than the control. Addition of anti-uPAR antibody in the cells remarkably down-regulated the phosphorylation level of p44 as compared with the control group, suggesting that uPA-ATF participates cell signal transduction and this reaction can be inhibited by anti-uPAR antibody; 5) uPA-ATF cell signal pathway up-regulated expression of MMP-2 and TIMP-3, while anti-uPAR antibody down-regulated the expression of MMP-2 and TIMP-3. CONCLUSION: These results demonstrate for the first time that uPA-ATF directly regulates the expression of MMP-2 and TIMP-3 by signal transduction pathway, and the over-expression of MMP-2 and TIMP-3 may play an important role in local osteolysis of GCT.     

Astrocyte proliferation in the rat hippocampus after middle cerebral artery occlusion

AIM: To study rat astrocyte proliferation in ipsilateral hippocampus following focal cerebral ischemia. METHODS: Ischemia was induced by temporary middle cerebral artery occlusion (MCAO). In hippocampus of rats at 3, 7 and 30 days after MCAO, the numbers and anatomic distribution of glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry. The protein expression of GFAP and proliferating cell nuclear antigen (PCNA) in the ipsilateral hippocampus were analyzed by Western blot analysis. RESULTS: Astrocytes appeared hypertrophic, with increased process thickness and numbers at 7 days after MCAO, and the highest density of astrocytes were seen at 30 days in the CA1, CA2 regions of the ipsilateral hippocampus. Western blot analysis revealed that GFAP levels were normal at 3 days, but increased by 7 days and remained elevation at 30 days. Western blot analysis of PCNA protein also revealed identified upregulation PCNA at 3 days after MCAO and the expression peaked at 7 days. CONCLUSION: This study demonstrates that focal cerebral ischemia in the rat results in a rapid response, a process often referred to as reactive astrogliosis or glial scarring, from resident astrocytes of the ipsilateral hippocampus to the side of ischemia.     

Effects of β-ME and RA on expression of GFAP in mesenchymal cells derived from mouse fetal liver

AIM: To investigate the effects of β-mercaptoethanol (β-ME) and all-trans rentinal acid (RA) on glial fibrillary acidic protein (GFAP) expression in mesenchymal cells derived from mouse fetal liver in vitro. METHODS: Cells suspension from 14.5-days-old mouse fetal liver were cultured in DMEM/HEPES/F12 supplemented with 20％ FCS and mesenchymal cells were acquired after discarding nonadherent cells. The 5th passage cells were induced by β-ME and RA. The characteristics of treated cells were assayed by immunocytochemistry staining at 5 hours and 5 days after induction. β-actin as an internal control, GFAP gene expression of mesenchyal cells was detected with semi-quantitative RT-PCR. RESULTS: After being inducted by β-ME and RA, 80％ approximately of the cells exhibited typical neural morphology and about 85％ expressed GFAP phenotype. Semi-quantitative RT-PCR showed that mRNA expression of GFAP increased in treated cells versus untreated cells (P<0.01). CONCLUSION: GFAP expression in mesenchymal cells derived from mouse fetal liver in vitro increases after being treated with β-ME and RA.     

Angiopoietin-1 and regulation of vascular permeability

Angiopoietin-1 (Ang-1) is a newly-found endothelium-specific proangiogenic factor and it had been proved essential roles in both vasculogenesis and angiogensis. Among them, its anti-leakage ability may have great potential applications in clinical treatment of vascular hyper-permeability in a variety of diseases such as cancer, diabetic retinopathy, rheumatoid arthritis, asthma. In this review, some research progresses focused on this aspect are discussed.     

禽大肠杆菌外膜蛋白基因C(ompC)的序列分析

根据 Gen Bank中人源大肠杆菌 K- 12外膜蛋白基因 C(omp C)的核苷酸序列设计引物 ,应用 PCR方法从禽大肠杆菌 O2 、O78株及它们的融合双价弱毒菌株 O2 ,78(Norr Chlr)中分别扩增得到 omp C基因 ,序列测定及分析比较发现 ,3个菌株的 om p C基因均由 170 2 nt组成 ,核苷酸序列完全相同 ,只有 1个大的开放性阅读框 (ORF) ,长 10 92 bp,编码由 36 3个氨基酸组成的前 Om p C蛋白 ,前 2 1个氨基酸残基组成信号肽 ,成熟的 Omp C蛋白由 342个氨基酸残基组成 ,Mr为 4 0 0 0 0。其氨基酸序列也完全相同。从基因水平上证明了禽大肠杆菌 O2 、O78株及融合双价弱毒菌株 O2 ,78(NorrChlr)存在相同的外膜蛋白 C抗原 ,从而为进一步研究 Omp C蛋白的免疫原性奠定了基础